Rapid On-Chip Isolation of Cancer-Associated Exosomes and Combined Analysis of Exosomes and Exosomal Proteins.
Rapid On-Chip Isolation of Cancer-Associated Exosomes and Combined Analysis of Exosomes and Exosomal Proteins.
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DOI：
10.1021/acs.analchem.2c01187
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发表时间：
2022-05
影响因子：
7.4
通讯作者：
Lu Zheng;Hua Wang;P. Zuo;Yuelin Liu;Huiying Xu;B. Ye
中科院分区：
文献类型：
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作者：
Lu Zheng;Hua Wang;P. Zuo;Yuelin Liu;Huiying Xu;B. Ye
胞外体是由多种细胞分泌的脂类双层胞外囊泡，从亲本细胞中继承了丰富的分子信息。肿瘤来源的外切体已被广泛认为是癌症早期诊断和监测的非侵入性生物标志物，但完整外切体的分离和外切体蛋白的检测仍然具有挑战性。在这里，我们提出了一种微流控芯片，用于特异性外切体的分离，并通过一种新的PTCDI-适配子信号开关策略集成了灵敏的定量。为了提高捕获效率，设计了一种交替的滴状微柱阵列，以帮助芯片上的Tim4修饰磁珠捕获肿瘤来源的外切体。捕获后，螯合剂可以很容易地洗脱完整的外体，这些外体通过多重荧光开启方法进一步用于分析外体表面蛋白。得益于高效的Tim4小球在芯片上的富集性和优越的荧光信号转导策略，该分析平台对HepG2外切体的检测下限低至8.69×103颗粒/毫升，线性范围宽，范围可达6个数量级。同时，所提出的平台可以识别来自不同细胞系的外体表面蛋白质水平的细微变化。更重要的是，这一策略被成功地应用于分析人血清中的外切体，以区分肝癌患者和健康人。联合分析外体膜表面不同类型的生物标志物可以极大地提高癌症类型识别和疾病监测的准确性。我们希望这个方便、快速、灵敏的平台可以成为外显体分析和癌症早期筛查领域的有力工具。
Exosomes are lipid bilayer extracellular vesicles secreted by various types of cells and inherit abundant molecular information from parental cells. Tumor-derived exosomes have been widely recognized as noninvasive biomarkers for early cancer diagnosis and surveillance, but the separation of intact exosomes and detection of exosomal proteins remain challenging. Herein, we proposed a microfluidic chip for specific exosome isolation, integrated with sensitive quantification by a novel PTCDI-aptamer signal switch strategy. To enhance the capture efficiency, an alternating drop-shaped micropillar array was designed to assist the capture of tumor-derived exosomes by Tim4-modified magnetic beads (Tim4 beads) on the chip. Following capture, a chelating agent can easily elute intact exosomes which were further used for profiling exosomal surface proteins by the multiplexed fluorescence turn-on approach. Profiting from the efficient on-chip enrichment of the Tim4 beads and superior fluorescence signal transduction strategy, the detection limit of the analysis platform for HepG2 exosomes is as low as 8.69 × 103 particles/mL with a wide linear range spanning 6 orders of magnitude. Meanwhile, the proposed platform could recognize subtle changes in protein levels on the exosomal surface from various cell lines. More importantly, this strategy is successfully applied to analyze exosomes in human serum to distinguish liver cancer patients from healthy individuals. Combined analysis of different types of biomarkers on the exosomal membrane surface can greatly improve the accuracy of cancer type identification and disease monitoring. We hope that this convenient, rapid, and sensitive platform may become a powerful tool in the field of exosome analysis and early cancer screening.